%0 Journal Article
%T GENE CLONING AND EXPRESSION OF PROLYL ENDOPEPTIDASE FROM AEROMONAS PUNCTA TA<br>点状产气单胞菌脯氨酰内肽酶基因的克隆与表达
%A Li Min
%A Chen Changqing
%A Wang Debao
%A <br>李民
%A 陈常庆
%A 王德宝
%J 微生物学报
%D 2000
%I 
%X Prolyl endopeptidase gene was cloned from Aeromonas punctata subsp. Punctata(ST-78-3-3) using activity screening method and the 3.3 kb DNA fragment containing PEP gene was sequenced. DNA sequence from 20-2092 bp was ORF region coding 690 amino acids of prolyl endopeptidase. It was a new PEP gene through gene search. The genetic engineered E. coli BL21/pGEM-PEP overexpressing recombinant Aeromonas punctata prolyl endopeptidase(apPEP) was constructed. Cultured in YH medium, the expressed apPEP was about 30% of total cellular protein, the activity was 112 times more than that of wild strain. Expressed apPEP was mainly soluble intracellular protein, about 5% of apPEP was secreted to medium. Non-reduced SDS-PAGE analysis showed it's monomer with molecular weight about 76 kD, which corresponded with prediction by gene sequence. Recombinant apPEP was purified after tube culture, the purity reached 90% and specific activity was 67 U/mg.
%K Prolyl endopeptidase
%K E
%K coli
%K Cloning
%K Overexpression<br>脯氨酰内肽酶，
%K 大肠杆菌，
%K 克隆，
%K 高效表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=8F5B905C4A58D26A&yid=9806D0D4EAA9BED3&vid=1371F55DA51B6E64&iid=38B194292C032A66&sid=CAA7BAE04CB631A1&eid=E2B9962CCD971A0D&journal_id=0001-6209&journal_name=微生物学报&referenced_num=2&reference_num=19